New anti-inflammatory and non-cytotoxic metabolites of methylstenbolone obtained by microbial transformation

• Published in: Bioorganic chemistry 2022-12, Vol.129, p.106187, Article 106187
• Main Authors: Aamer, Muhammad, Siddiqui, Mahwish, Jabeen, Almas, Irshad, Rimsha, Khan, Farooq-Ahmad, Atia-tul-Wahab, Iqbal Choudhary, M., Wang, Yan
• Format: Article
• Language: English
• Subjects: Anabolic steroid; Androstenols; Anti-inflammatory activity; Anti-Inflammatory Agents - pharmacology; Biotransformation; Cunninghamella blakesleeana; Cytotoxicity; Humans; Macrophomina phaseolina; Methylstenbolone; NO production
• ISSN: 0045-2068; 1090-2120
• DOI: 10.1016/j.bioorg.2022.106187

[Display omitted] •Methylstenbolone was transformed by fungi for the first time.•Nine transformed analogues were reported, eight of which are new.•Methylstenbolone was discovered as a potent anti-inflammatory agent.•Some analogues showed good anti-inflammatory activity.•Methylstenbolone is cytotoxic, but metabolites were not. A synthetic anabolic–androgenic steroid, methylstenbolone (1), was structurally transformed into a series of nine analogues, 2,17α-dimethyl-7α,17β-dihydroxy-5α-androst-1-en-3-one (2), 2,17α-dimethyl-15β,17β-dihydroxy-5α-androst-1-en-3-one (3), 2,17α-dimethyl-6α,9α,17β-trihydroxy-5α-androst-1-en-3-one (4), 2-methyl-17β-hydroxy-17α-(hydroxymethyl)-5α-androst-1-en-3-one (5), 2-methyl-11β,17β-dihydroxy-17α-(hydroxymethyl)-5α-androst-1-en-3-one (6), 2-methyl-17β-hydroxy-17α-(hydroxymethyl)-5α-androst-1-en-3,6-dione (7), 2-methyl-17β-hydroxy-17α-(hydroxymethyl)-5β-androst-1-en-3,6-dione (8), 2,17α-dimethyl-7β,17β-dihydroxy-5α-androst-1-en-3-one (9), and 2,17α-dimethyl-12β,17β-hydroxy-5α-androst-1-en-3,7-dione (10) by fungal cell suspension cultures, Macrophomina phaseolina and Cunninghamella blakesleeana for the first time. Among those, compounds 2–4 and 6–10 were identified as new. Herein, spectral data of metabolite 5 was reported for the first time. Their structures were elucidated by NMR, MS, UV, and IR spectroscopic methods. Substrate 1 (IC50 10.1 ± 0.3 µg/mL) was identified as a potent anti-inflammatory agent against nitric oxide (NO) production. Its transformed products 3 (IC50 as 27.8 ± 1.1 µg/mL) and 9 (26.9 ± 0.4 µg/mL) displayed good inhibition. Compounds 2 (IC50 = 45.9 ± 0.8 µg/mL) and 6 (IC50 = 36.6 ± 1.2 µg/mL) were also active moderately against NO production, in comparison to standard LNMMA (IC50 = 24.2 ± 0.8 µg/mL). Cytotoxicity assay showed 1 was active to cancer cell line MCF7 (IC50 = 12.26 ± 0.35 µg/mL), compared to the standard Doxorubicin having IC50 as 1.25 ± 0.11 µg/mL. However, it is also toxic to human normal cell line (BJ) with IC50 as 8.69 ± 0.02 µg/mL. More importantly, all transformed products are non-cytotoxic on BJ. Therefore, biotransformation can be an efficient approach to reduce the toxicity of methylstenbolone.